Radioimmunoprecipitation Assay for Australia Antibody

2015 ◽  
pp. 129-136
Author(s):  
A. Schober ◽  
R. Thomssen
Keyword(s):  
Radioimmunoprecipitation Assay

Radioimmunoprecipitation Assay for Australia Antibody

Vox Sanguinis ◽  
1973 ◽  
Vol 24 (1) ◽  
pp. 58-65
Author(s):  
A. Schober ◽  
R. Thomssen
Keyword(s):  
Radioimmunoprecipitation Assay

Radioimmunoprecipitation Assay for Australia Antigen, Antibody, and Antigen-Antibody Complexes

1971 ◽  
Vol 138 (1) ◽  
pp. 249-257 ◽  
Author(s):  
J. A. Coller ◽  
I. Millman ◽  
T. C. Halbherr ◽  
B. S. Blumberg
Keyword(s):  
Australia Antigen ◽  
Radioimmunoprecipitation Assay ◽  
Antigen Antibody

scholarly journals Antibodies in human sera to oncorna virus-like proteins from normal or leukemia marrow cell cultures.

1976 ◽  
Vol 144 (5) ◽  
pp. 1243-1253 ◽  
Author(s):  
S Louie ◽  
J E Curtis ◽  
J E Till ◽  
E A McCulloch
Keyword(s):  
Lupus Erythematosus ◽  
Competitive Inhibition ◽  
Marrow Cell ◽  
Leukemia Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cross Reactivity ◽  
Antibody Activity ◽  
Human Sera ◽  
Blood Specimens ◽  
Radioimmunoprecipitation Assay

Some human marrows in culture release particles with oncornavirus-like properties. This study was designed to examine the immunological properties of similar particles in human marrow culture supernates. Leukemic and nonleukemic marrows were cultured for 5-7 days in the presence of [14C]uridine and [3H]leucine or [3H]glucosamine. Labeled supernatant components banding in sucrose gradient densities of 1.20-1.24 g/ml were used as antigen in a double antibody immunoprecipitation assay. The assay was validated by end point titrations and competition with unlabeled antigen; purified myeloma proteins were used as negative controls. Cross-reactivity with mammalian oncornaviruses, as judged by competitive inhibition of precipitation by these viruses, was slight and at the border of the sensitivity of the method. Precipitated antigens analyzed by SDS polyacrylamide gel electrophoresis contained three distinct polypeptides of about 70,000, 45,000 and 30,000 mol wt; these comigrated with the gp 70, pg 45, and p 30 of a murine leukemia virus. Similar polypeptides were obtained from both leukemic and nonleukemic marrow culture supernates. As determined by the radioimmunoprecipitation assay, 32 of 45 leukemic sera (71%), 36 of 45 normal sera (80%), 15 of 19 sera from family contacts of leukemic patients (79%), 14 of 21 cord blood specimens (67%), and 21 of 23 sera (91%) from patients with systemic lupus erythematosus had detectable antibody activity.

Autoimmune Peripheral Nervous System Hyperexcitability

2021 ◽  
pp. 138-139
Author(s):  
Christopher J. Klein
Keyword(s):  
Potassium Channel ◽  
Intravenous Immunoglobulin ◽  
Immunoglobulin G ◽  
High Dose ◽  
Voltage Gated ◽  
Idiopathic Pain ◽  
Upper And Lower Extremities ◽  
History Of ◽  
Reflex Testing ◽  
Radioimmunoprecipitation Assay

A 25-year-old man was seen for assessment of progressive pain. He had a distant history of Guillain-Barré syndrome at age 8 years, at which time he had symmetrical proximal and distal weakness of the upper and lower extremities with loss of ambulation. No facial weakness, dysarthria, dysphagia, ptosis, diplopia, or respiratory weakness occurred. At his initial evaluation there was touch hypersensitivity of the muscles and skin. He had no weakness or cognitive involvement, although the pain made it difficult for him to concentrate. His creatine kinase value improved with hydration, but pain and muscle twitching persisted. On examination, he had diffuse extremity and truncal fasciculations and myokymia and reported pain in not only the areas of twitching but also other areas of his extremities and trunk. On neurophysiologic testing, fibular and tibial motor compound muscle action potentials were decreased in amplitude, with normal ulnar and median motor responses. Needle electromyography of muscles proximally and distally showed diffuse spontaneous firing of muscles ranging in frequency with waxing and waning characteristics. These findings were thought to be consistent with a primary hyperexcitable disorder of muscles with a superimposed old polyradiculoneuropathy and possibly a myopathy. Expanded autoimmune neuroimmunologic testing of serum identified immunoglobulin G-directed cerebellar molecular staining consistent with voltage-gated potassium channel autoantibodies. Radioimmunoprecipitation assay identified voltage-gated potassium channel-immunoglobulin Gs and led to reflex testing for contactin-associated protein 2-immunoglobulin G; autoantibodies were positive. Computed tomography of the chest with contrast was performed, and lymphadenopathy was identified. The patient was clinically diagnosed with contactin-associated protein 2 - immunoglobulin G–positive Isaacs syndrome. A trial of high-dose gabapentin was attempted, with only mild benefits. Next, intravenous immunoglobulin was initiated. Diabetes developed, and he was hospitalized requiring initiation of insulin. His condition is now managed variably with intravenous immunoglobulin and scheduled daily gabapentin. The immune system has long been recognized to help regulate pain via non- immunoglobulin G–mediated mechanisms. Specifically, cytokines decrease the nociceptive nerve fiber thresholds and are released after diverse tissue insults. This allows for speeded healing by increased blood flow and protection of the region by pain guarding mechanisms. It is now recognized that, in rare cases, immunoglobulin G-mediated autoimmunity can lead to otherwise idiopathic pain disorders.

scholarly journals A RADIOIMMUNOPRECIPITATION ASSAY (RIPA) FOR ANTIBODIES TO GROWTH HORMONE (GH)

1988 ◽  
Vol 23 (1) ◽  
pp. 118-118
Author(s):  
L Di Silvio ◽  
P Hindmarsh ◽  
J Pringle ◽  
C Brook ◽  
A Kurtz
Keyword(s):  
Growth Hormone ◽  
Radioimmunoprecipitation Assay

Monitoring ADAMTS13 Autoantibody Titer Using a Novel Quantitative Radioimmunoprecipitation Assay In Patients With Acquired Thrombotic Thrombocytopenic Purpura

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3561-3561
Author(s):  
Atsuko Igari ◽  
Takanori Moriki ◽  
Hideo Wada ◽  
Kenji Soejima ◽  
Mitsuru Murata
Keyword(s):  
Monoclonal Antibodies ◽  
Plasma Exchange ◽  
Thrombotic Thrombocytopenic Purpura ◽  
Conflicts Of Interest ◽  
Expression System ◽  
Plasma Samples ◽  
Thrombocytopenic Purpura ◽  
Free Protein ◽  
Autoantibody Titer ◽  
Radioimmunoprecipitation Assay

Abstract Background ADAMTS13-binding immunoglobulin G (IgG)-type autoantibodies are present in patients with acquired thrombotic thrombocytopenic purpura (TTP), thereby causing severe deficiency of plasma ADAMTS13 activity. Some specific autoantibodies directly inhibit the enzymatic activity by interfering with the access of its substrate, von Willebrand factor (VWF), particularly in the spacer domain, although others bind to almost all of the domains in the molecule, possibly leading to the acceleration of the clearance from blood stream. Not only the inhibitor titer, but the importance of ADAMTS13 autoantibody titer was highlighted by many previous clinical studies, associating with the prognosis such as recurrence rate. Objective We targeted to establish a novel high-sensitive assay to measure ADAMTS13-binding IgG autoantibody titer using three kinds of radioisotope-labeled antigens, ADAMTS13 whole molecule, MDTCS and T2-8/CUB, and aimed to analyze the association between the autoantibody titer and the clinical characteristics of TTP patients. Materials and Methods Human Cell-Free Protein Expression System (Takara #3281, Shiga, Japan) was used to synthesize radioisotope-labeled antigens. ADAMTS13 cDNA corresponding to whole molecule (A13), metalloprotease to spacer domains (MDTCS) and TSP1-2 to CUB2 domains (T2-8/CUB) were cloned into an expression vector pT7-IRES and mixed with Cell Lysate containing T7 RNA polymerase, methionine-free amino acids, ATP, translation enhancement factor and 35S-methionine. The correct synthesis and molecular size of the radiolabeled antigens were checked with SDS-PAGE. To assess the utility as a quantitative assay, each of the antigens was mixed with mouse anti-ADAMTS13 monoclonal antibodies, whose epitopes were determined in our previous study (Thromb Res. 2012; 130(3):e79-83), and the immune complex was precipitated with protein G beads, washed and measured in a liquid scintillation counter. Plasma samples from acquired TTP patients were tested to quantify the autoantibody titers using radiolabeled A13, MDTCS and T2-8/CUB antigens, respectively. As a control, plasma samples from healthy subjects with no histories of autoimmune disease were also tested. Results Each of the radiolabeled antigens was detected as a single band at the correct molecular weight size and successfully immnoprecipitated with several mouse anti-ADAMTS13 monoclonal antibodies, indicating the intact molecular conformation of the synthesized proteins using the cell-free protein synthesis system. Moreover, the appropriate dose-dependent escalation curves in accordance with the addition of the monoclonal antibodies were observed, thereby confirming the utility of the assay as a quantitative analysis. We tested TTP patient plasma at onset (n=5) and were able to detect ADAMTS13 autoantibody titers with each of the radiolabeled A13, MDTCS and T2-8/CUB antigens. We next applied this assay for monitoring ADAMTS13 autoantibody titer in a clinical course of TTP patient. The patient developed the first episode of TTP at the age of 2 month and treated with steroid pulse and plasma exchange therapy for six consecutive days. Remission was once achieved but 6 months later from the onset, the second episode of TTP occurred and the patient was treated with 11 plasma exchange and rituximab at the dose of 375 mg/m2 once a week for 4 weeks. Plasma samples at onset, after the first 6 consecutive plasma exchange and after rituximab administration were examined about autoantibody titer using this assay. Interestingly, the titers remained high even after the plasma exchange but declined clearly after the rituximab treatment, whereas no reduction of total IgG level was observed. These findings suggest that the autoantibody titration using this assay might be useful to assess the effect of treatment and associate with the prognosis related to the recurrence. Conclusion We developed a novel quantitative radioimmunoprecipitation assay to measure ADAMTS13 autoantibody titer related to three antigens, ADAMTS13 whole molecule, MDTCS and TSP2-8/CUB. This assay may serve not only as a diagnostic test but as a monitoring index to evaluate the prognosis of TTP. Disclosures: No relevant conflicts of interest to declare.

Clinical Evaluation of a Radioimmunoprecipitation Assay for IA-2 Antibody and Comparison of GAD Antibody in Type 1 Diabetes Mellitus

Autoimmunity ◽  
2000 ◽  
Vol 32 (2) ◽  
pp. 79-88 ◽  
Author(s):  
Mitsuhiro Ohta ◽  
Kiyoe Ohta ◽  
Hiroshi Obayashi ◽  
Naoto Nakamura ◽  
Hirofumi Shlgeta ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Type 1 Diabetes ◽  
Type 1 Diabetes Mellitus ◽  
Clinical Evaluation ◽  
Radioimmunoprecipitation Assay
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